| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| Other Sizes |
| Targets |
This peptide fragment acts as a ligand for Notch receptors, but its primary utility lies in studying the interactions and activation of the Notch signaling pathway. Upon binding to its physiological ligands (such as Jagged1 and Delta-like family members), Notch receptors undergo proteolytic cleavage by ADAM-family proteases and the gamma-secretase complex, leading to the release of the Notch intracellular domain (NICD). The NICD then translocates to the nucleus, where it forms a transcriptional activation complex with the DNA-binding protein CSL (CBF1/Su(H)/Lag-1) and co-activators like MAML, ultimately driving the expression of target genes such as Hes1, Hey1, and c-Myc. The Notch 1 peptide is essential for exploring these fundamental receptor-ligand interactions and the initiation of signaling.
|
|---|---|
| ln Vitro |
It has been demonstrated that Notch 1 increases the survival and proliferation of mouse luminal cells by activating the prosurvival NF-κB pathway[2]. In vitro invasion and cell proliferation are reduced when Notch 1 and Jagged1 are downregulated in human prostate cancer cell lines[2].
In vitro studies have shown that Notch signaling promotes the survival and proliferation of mouse luminal prostate cells by activating the pro-survival NF-kappaB pathway. This pathway's activation is mediated through Notch1, which can upregulate components of the NF-kappaB signaling cascade. Conversely, the down-regulation of Notch1 and its ligand Jagged1 in human prostate cancer cell lines has been demonstrated to significantly decrease cell invasion and growth. Furthermore, Notch1 signaling can influence epithelial-to-mesenchymal transition (EMT), contributing to the invasive and metastatic potential of cancer cells. This highlights the dual role of Notch in both normal development and oncogenesis. |
| ln Vivo |
No specific in vivo data is available for this exact Notch 1 peptide fragment. However, the in vivo role of the full-length Notch1 receptor has been extensively characterized. Notch1 is critically involved in T-cell development and the maintenance of the intestinal epithelium. Aberrant activation of Notch1 signaling in murine models has been linked to the development of T-cell acute lymphoblastic leukemia (T-ALL). Furthermore, the use of gamma-secretase inhibitors (GSIs) or anti-Notch1 antibodies in xenograft models has demonstrated anti-tumor activity by blocking Notch signaling. The systemic inhibition of Notch1 in vivo can cause significant intestinal goblet cell metaplasia, highlighting the importance of Notch in gut homeostasis.
|
| Enzyme Assay |
Cell-free binding assays are typically performed using Surface Plasmon Resonance (SPR) or Enzyme-Linked Immunosorbent Assays (ELISA) to measure the interaction between the Notch1 fragment and its ligands or antibodies. For an ELISA, a 96-well plate is coated with the Notch 1 peptide (1-5 microg/mL) in coating buffer overnight at 4degC. The plate is then blocked with 3% BSA or 5% non-fat milk in PBS-Tween (PBST) for 1 hour at room temperature. Serially diluted recombinant Jagged1 or an anti-Notch1 antibody is added and incubated for 2 hours. After washing, a secondary HRP-conjugated antibody is added, followed by TMB substrate. The reaction is stopped with H2SO4, and absorbance is read at 450 nm to calculate binding affinity (Kd) or to assess the blocking ability of competitor molecules. This format allows for high-throughput screening of Notch1-binding compounds.
|
| Cell Assay |
A typical protocol involves the use of human prostate cancer cell lines such as PC-3, DU145, or LNCaP. Cells are seeded in 6-well plates at 70-80% confluency and allowed to attach overnight. For Notch1 down-regulation, cells are transfected with 50-100 nM of siRNA targeting Notch1 or Jagged1 using a lipid-based transfection reagent (e.g., Lipofectamine 2000). After 48-72 hours, cells are harvested for analysis. Cell proliferation is assessed by MTT or CellTiter-Glo assays according to the manufacturer's instructions. For invasion assays, 1×10^5 cells in serum-free medium are seeded into Matrigel-coated transwell inserts. The lower chamber contains medium with 10% FBS as a chemoattractant. After 24-48 hours, non-invading cells are removed from the upper chamber with a cotton swab, and invading cells are stained with crystal violet, then quantified by microscopic counting or eluting the stain for absorbance measurement at 590 nm.
|
| Animal Protocol |
The Notch signaling pathway is frequently studied in vivo using xenograft mouse models, such as C57BL/6 or athymic nude mice bearing human tumor xenografts. For anti-tumor efficacy studies, 5×10^6 prostate cancer cells are injected subcutaneously into the flank of the mouse. Once tumors reach an average volume of 100-200 mm3, treatment with a Notch pathway inhibitor is initiated. A common regimen is intraperitoneal (IP) administration of a gamma-secretase inhibitor (e.g., 5-10 mg/kg) or a blocking antibody daily for 2-3 weeks. Tumor volumes are measured with calipers every 2-3 days and calculated using the formula: volume = (length × width2)/2. At the end of the study, tumors are excised, and NICD levels are quantified in tumor lysates via Western blotting to confirm target engagement. Immunohistochemistry (IHC) for Ki-67 can be used to assess tumor cell proliferation.
|
| ADME/Pharmacokinetics |
The lyophilized powder is stable for up to 3 years when stored at -20degC, protected from moisture. For long-term storage of solutions, it is recommended to store at -80degC for up to 6 months, avoiding multiple freeze-thaw cycles to prevent peptide degradation. The compound is soluble in water at approximately 18.5 mg/mL, making it suitable for aqueous-based assays. For in vivo formulation, it can be dissolved in a mixture of DMSO, Tween 80, and saline (e.g., 10:5:85 ratio) to achieve a clear solution appropriate for injection. The peptide has a molecular weight of 1614.81 g/mol and its purity is typically greater than 95%. Because it is a peptide, it is likely subject to rapid proteolytic degradation and clearance in serum, resulting in a short plasma half-life of minutes to hours.
|
| Toxicity/Toxicokinetics |
As a research-grade chemical, Notch 1 TFA is not intended for human therapeutic use and has no defined toxicity profile. The primary toxicity concerns relate to the TFA counterion, which can be cytotoxic to some sensitive cell types if present in high concentrations. It is recommended to keep the TFA concentration below 0.1% in cell culture media to avoid non-specific effects. In animal studies, inhibitors of the Notch pathway are known to cause significant on-target toxicities, most notably severe gastrointestinal toxicity characterized by goblet cell hyperplasia and villus atrophy, as Notch1 is essential for maintaining the intestinal stem cell compartment. General safety precautions for laboratory handling include the use of appropriate personal protective equipment (PPE) to avoid inhalation, ingestion, or skin contact.
|
| References | |
| Additional Infomation |
This product is strictly for research use only and is not approved for clinical applications or human use. The trifluoroacetic acid (TFA) salt form enhances the peptide's solubility and stability compared to the free base form. However, researchers should be aware that residual TFA may interfere with certain cell-based assays or biological systems. The Notch family consists of four receptors (Notch1-4) and five canonical ligands (Jagged1, Jagged2, Dll1, Dll3, Dll4). Dysregulation of Notch signaling is a hallmark of many human cancers, including T-ALL, breast cancer, and colorectal cancer, making it a significant target for anti-cancer drug discovery. Pharmacological modulators include gamma-secretase inhibitors and blocking antibodies, but as a peptide fragment, this product serves as a valuable tool for molecular interaction studies and screening assays.
|
| Molecular Formula |
C64H98N15F3O25S3
|
|---|---|
| Molecular Weight |
1614.81
|
| Appearance |
Off-white to light yellow solid powder
|
| HS Tariff Code |
2934.99.9001
|
| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
H2O :~18.52 mg/mL (~11.47 mM)
|
|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.6193 mL | 3.0963 mL | 6.1927 mL | |
| 5 mM | 0.1239 mL | 0.6193 mL | 1.2385 mL | |
| 10 mM | 0.0619 mL | 0.3096 mL | 0.6193 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
